GENE THERAPY OF GLOMERULONEPHRITIS BY GENE TRANSFER OF CELL CYCLE-RELATED GENES.

通过细胞周期相关基因的基因转移进行肾小球肾炎的基因治疗。

• 批准号: 09557090
• 负责人: TERADA Yoshio
• 金额: $ 7.55万
• 依托单位: TOKYO MEDICAL AND DENTAL UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-09557090/
• 关键词: Adenovirus; cyclin; cell cycle; Glomerulonephritis; mesangial cell; cGMP; HMG-Coa inhibitor; glucocorticoid; 腎炎; メザンギウム細胞; アデノカイルス

Recent studies have revealed that mitogen-actiated protein kinase(MAPK)consists of at least three subfamilies, namely include classical MAPK(also known as ERK), stress-activated protein kinase/c-Jun N-terminal kinase(JNK), and p38 kinase. TGF-b-activating kinase(TAK)-1 is a novel MAPKKK which is reported to stimulate p38K and/or JNK pathway. To elucidate functional roles of the TAK1 pathway, we transfected its constitutive active form(TAKdN)and negative form(TAKK63W)to LLCPK1 cells. TAKdN inhibited 3H-thymidine uptake, and reduced the percentages of S and G2/M phases. TAKK63W ameliorated inhibition of 3H-thymidine uptake and the percentages of S and G2/M phases by TGF-b. Western blot analysis demonstrates that the cyclin D1 protein level was negatively regulated by overexpression of TAKdN.Moreover, overexpression of TAKdN inhibited cyclin D1 promoter activity. In contract, constitutive active MKK1, the classical p42/44 MAPK-activator, increased cyclin D1 promoter activity and protein level. Overexpression of the active form of MKK1 increased 3H-thymidine uptake, while the inactive form decreased the uptake.In conclusion, cyclin D1 promoter activity and cell cycle progression are negatively regulated by the TAK1-MKK6-p38K pathway and positively regulated by the MKK1-p42/44 MAPK pathway.

最近的研究表明，有丝分裂原活性蛋白激酶（MAPK）由至少三个亚家族组成，即包括经典的MAPK（也称为ERK），应激激活的蛋白激酶/C-JUN N末端激酶（JNK）和P38激酶。 TGF-B激活激酶（TAK）-1是一种新型的MAPKKK，据报道刺激P38K和/或JNK途径。为了阐明TAK1途径的功能作用，我们将其本构的活动形式（TAKDN）和负形式（TAKK63W）转染至LLCPK1细胞。 TAKDN抑制了3H-胸腺定氨酸的摄取，并降低了S和G2/M相的百分比。 TAKK63W改善了TGF-B的3H-胸苷摄取的抑制作用以及S和G2/M相的百分比。 Western印迹分析表明，Cyclin D1蛋白水平受TAKDN的过表达对负调节。此外，TAKDN抑制细胞周期蛋白D1启动子活性的过表达。在合同中，经典P42/44 MAPK-激活剂的组成型Active MKK1增加了细胞周期蛋白D1启动子活性和蛋白质水平。 MKK1活性形式的过表达增加了3H-胸腺素的摄取，而无活性形式降低了摄取。结论，Cyclin D1启动子活性和细胞周期进程受到TAK1-MKK6-P38K途径的负调节，并由MKK1-P42/44 MAPK途径阳性调节。

项目成果

Y.Terada, T.Yamada, O.Nakashima, S.Sasaki, H.Nonoguchi, K.Tomita, F.Marumo: "Expression of PDGF and PDGF receptor mRNA in glomerulus of IgA nephropathy."Journal of American Society of Nephrology. 8. 817-819 (1997)

Y.Terada、T.Yamada、O.Nakashima、S.Sasaki、H.Nonoguchi、K.Tomita、F.Marumo：“IgA 肾病肾小球中 PDGF 和 PDGF 受体 mRNA 的表达”。美国肾脏病学会杂志。

Y.Terada et al: "Overexpression of cell cycle inbibitors (p161NK4 and p21 cipl) and cyclin D1 using adenovirus regulates proliferation of rat mesangial cells." Journal of American Society of Nephrology. 8. 51-60 (1997)

Y.Terada 等人：“使用腺病毒过度表达细胞周期抑制剂（p161NK4 和 p21 cipl）和细胞周期蛋白 D1 可调节大鼠系膜细胞的增殖。”

Y. Terada, O. Nakashima, S. Inoshita, M. Kuwahara, S. Sasaki, F. Marumo.: "TGF-β-activating kinase-1 inhibits cell cycle and expression of cyclin D1 and A in LLC-PK1 cells."Kidney International. 56. 1378-1390 (1999)

Y. Terada、O. Nakashima、S. Inoshita、M. Kuwahara、S. Sasaki、F. Marumo.：“TGF-β 激活激酶-1 抑制 LLC-PK1 细胞中的细胞周期和细胞周期蛋白 D1 和 A 的表达。 “国际肾脏病杂志。56。1378-1390 (1999)

S.Inoshita et al: "Roles of E2F1 in mesangial cell cycle in vitro"Kidney International. 56. 2086-2095 (1999)

S.Inoshita 等人：“E2F1 在体外系膜细胞周期中的作用”Kidney International。

Y.Terada, T.Okado, S.Inoshita, S.Hanada, M.Kuwahara, S.Sasaki, T.Yamamoto, F.Marumo: "Glucocorticoids stimulate p21CIP1 and arrest cell cycle in vitro and in anti-GBM glomerulonephritis."Kidney International. (in press).

Y.Terada、T.Okado、S.Inoshita、S.Hanada、M.Kuwahara、S.Sasaki、T.Yamamoto、F.Marumo：“糖皮质激素在体外和抗 GBM 肾小球肾炎中刺激 p21CIP1 并阻止细胞周期。”